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picture1_Rflp Slideshare 68389 | Rflp Item Download 2022-08-28 19-10-24


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File: Rflp Slideshare 68389 | Rflp Item Download 2022-08-28 19-10-24
it is a method used by molecular biologists to follow a particular sequence of dna as it is passed on to other cells rflps can be used in many different ...

icon picture PPTX Filetype Power Point PPTX | Posted on 28 Aug 2022 | 3 years ago
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 It is a method used by molecular biologists to follow a 
  particular sequence of DNA as it is passed on to other cells. 
  RFLPs can be used in many different settings to accomplish 
  different objectives .It is a difference in homologous DNA 
  sequences that can be detected by the presence of fragments 
  of different lengths after digestion of the DNA samples in 
  question with specific restriction endonucleases. RFLP, as a 
  molecular marker, is specific to a single clone/restriction 
  enzyme combination. 
 Each organism inherits its DNA from its parents. Since DNA is 
  replicated with each generation, any given sequence can be 
  passed on to the next generation. An RFLP is a sequence of 
  DNA that has a restriction site on each end with a "target" 
  sequence in between. A target sequence is any segment of 
  DNA that bind to a probe by forming complementary base 
  pairs  that  hybridizes  with  one  or  more  fragments  of  the 
  digested  DNA  sample  after  they  were  separated  by  gel 
  electrophoresis,  thus  revealing  a  unique  blotting  pattern 
  characteristic to a specific genotype at a specific locus.
         RFLP probes are short, single- or low-copy genomic DNA 
  or cDNA clones are typically used. They are frequently used 
  in genome mapping and in variation analysis (genotyping, 
  forensics, paternity tests, hereditary disease diagnostics). Most 
  RFLP markers are co-dominant (both alleles in heterozygous 
  sample will be detected) and highly locus-specific. The RFLP 
  method is a direct test in which genomic DNA is isolated from 
  biological material and analyzed without any prior 
  amplification via the polymerase chain reaction (PCR). 
  
            The basic technique for detecting RFLPs involves 
  fragmenting a sample of DNA by a restriction enzyme, 
  which can recognize and cut DNA wherever 
  a specific short sequence occurs. The resulting DNA 
  fragments are then separated by length by agarose gel 
  electrophoresis, and transferred to a membrane via 
  the Southern blot procedure. Hybridization of the membrane 
  to a labeled DNA probe then determines the length of the 
  fragments which are complementary to the probe. 
 .
            An RFLP occurs when the length of a detected fragment 
  varies between individuals. Each fragment length is 
  considered an allele, and can be used in genetic analysis. 
  RFLP analysis may be subdivided into single- (SLP) and 
  multi-locus probe (MLP) paradigms. Usually, the SLP method 
  is preferred over MLP because it is more sensitive, easier to 
  interpret and capable of analyzing mixed-DNA samples 
  Briefly, DNA is extracted and cleaved into small fragments 
  using a selected restriction endonuclease and polymorphic 
  variation in the lengths of the resulting restriction fragments 
  is used to distinguish between individuals. 
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...It is a method used by molecular biologists to follow particular sequence of dna as passed on other cells rflps can be in many different settings accomplish objectives difference homologous sequences that detected the presence fragments lengths after digestion samples question with specific restriction endonucleases rflp marker single clone enzyme combination each organism inherits its from parents since replicated generation any given next an has site end target between segment bind probe forming complementary base pairs hybridizes one or more digested sample they were separated gel electrophoresis thus revealing unique blotting pattern characteristic genotype at locus probes are short low copy genomic cdna clones typically frequently genome mapping and variation analysis genotyping forensics paternity tests hereditary disease diagnostics most markers co dominant both alleles heterozygous will highly direct test which isolated biological material analyzed without prior amplification v...

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